TRAC CRM49: Making iPSC From Blood
Co-Sponsored by the NIH Center for Regenerative Medicine (CRM)
Recent advances in generating iPSCs now allow for their derivation from blood. This recent advance enables basic and clinical researchers to reprogram a blood cell into an iPSC and then further differentiate into any cell type. This capability allows researchers to develop "disease in a dish" paradigms to investigate disease and therapy mechanisms.
In this one week workshop, participants will learn how to generate iPSC from blood samples using a non-integrating approach. Due to the length of this procedure (iPSC generation ~3-4 weeks etc.) starting material (CD34+ cells or mononuclear cells) will be provided for each investigator and only critical stages will actually be performed during the laboratory portion of the workshop.
In addition to learning how to culture cells and reprogram blood cells into iPSCs, we will also present some of the latest methodologies for directing differentiation of these iPSCs into different lineages (e.g. hepatocytes or cardiomyocytes). Therefore, this course will package together the essential methodology to take a CD34+ cell isolated from blood, reprogram this cell, and then direct differentiation into multiple different lineages.
Lecture Topics: Introduction to Human Induced Pluripotent Stem Cells; Cellular Reprogramming Methodologies, Reprogramming from Blood; Specialized Media and Tools for Isolation and Enrichment of Blood-Derived Cells; Reprogramming using Cytotune Sendai Virus and Episomal Vectors; Differentiating IPSC to MSCs; Differentiating IPSC to RPEs; Overview of Validation Tests for Pluripotency; Discussion With Experts: Troubleshooting and Question Session, Planning Your Experiments the Right Way, Pam Robey, Ph.D (NIDCR), Rupa Shevde, Ph.D (Life Technologies), Barbara Mallon, Ph.D (Stem Cell Unit), Guokai Chen, Ph.D (NHLBI), Kapil Bharti, Ph.D (NEI).
Laboratory Topics: Observation of Monday’s cells and Cryopreservation; Maintenance of human iPSCs and Differentiation of iPSCs via Embryoid Bodies or Monolayer Platforms; iPS colony identification from TeSR-E7 Reprogramming Experiments; Making Embryoid Bodies Using Aggrewell; Neural Rosette Identification and Scoring; Reprogramming Using Cytotune Virus; Progression of Colony Formation via Fixed Dishes; Picking of Reprogrammed Colonies; Neon Electroporation, Transfection; Validation Tests for Pluripotency; RT-QPCR Assay for iPSC Characterization
Benefits for Early Registering
- Register by March 1, 2014 to take advantage of the early registration discount ($995). The registration fee will increase to $1,095 on March 2, 2014
- Discounted Cell Lines for workshop attendees - ND2 (fibroblast Origin) and NCRM-5 (Blood Origin), $187.55 per line + S&H
- Class size will be limited to 20 (waiting list anticipated, 2013 summer offering filled to capacity)
- This workshop will be offered only once in 2014
Related Bio-Trac Courses:
Bio-Trac 18: Stem Cells
Bio-Trac 47: Human Pluripotent Stem Cells (hiiPSC); Differentiation Neural Lineages
Bio-Trac CRM 49: Making iPSCs from Blood
Bio-Trac CRM 54: Making Cardiomyocytes from iPSCs
Bio-Trac CRM 55: Engineering with CRISPR, TALENs, and ZFNs